The quantitative measurements of apoptotic status based on caspase 3/7 activation were examined using Muse® Caspase 3/7 Activation Kit, EMD Millipore Co. (Billerica, MA, USA), as described previously.15 (link) Briefly, after 24 h treatment with VCX, cells were trypsinized, centrifuged at 300×g for 5 min, and then resuspended in 1X Assay Buffer, followed by incubation with 5 µL of caspase 3/7 reagent working solution for 30 min at 37°C in 5% of CO2 incubator. After incubation, 150 µL of Muse® Caspase 7-ADD working solution was added and incubated at room temperature for 5 min in the dark. The percentage of caspase 3/7 positive cells was measured using Muse® Cell Analyzer, EMD Millipore Co. (Billerica, MA, USA).
Muse caspase 3 7 activation kit
Manufactured by Merck Group
Sourced in United States, Germany
The Muse® Caspase 3/7 Activation Kit is a laboratory assay designed to detect and quantify the activity of caspase-3 and caspase-7 enzymes in cell samples. Caspase-3 and caspase-7 are key mediators of apoptosis, a form of programmed cell death. The kit provides a fluorescence-based method to measure the levels of activated caspase-3 and caspase-7 in cells.
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2 protocols using muse caspase 3 7 activation kit
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Quantitative Apoptosis Assay using Caspase 3/7
2
Caspase-Dependent Apoptosis Evaluation
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To further confirm whether ME and WE induced caspase-dependent apoptosis, the Muse® caspase3/7 activation Kit (Merck-Millipore, Germany) was employed. Briefly, the cells (2 × 103 cells/well) were cultured in plates. Following incubation, 24 h treatment with the IC50 s and the controls was carried out. The cells were detached from the plate using 1X trypsin, which was deactivated in complete medium, followed by collection at 300 x g for 5 min using centrifugation. The samples were re-suspended in 50 μL of 1X Assay Buffer and 5 μL of Muse® Caspase-3/7 Reagent (Merck-Millipore, Germany). The samples were thoroughly mixed by gently pipetting up and down for 5 s. The tube caps were loosened and incubated samples for 30 min in the 37 °C chamber. After incubation, the Muse® Caspase 7-AAD (Merck-Millipore, Germany) [150 μL] was added to each tube and placed in the dark at 25 °C for 5 min. The Muse® Cell Analyser (Merck-Millipore, Germany) was used to analyse the samples.
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